An Automated High-Throughput Screening Enzyme Linked Immunosorbent Assay for Johne's Disease Antibodies in Bovine Serum
Notice bibliographique
Résumé
Amanual Mycobacterium avium subspecies paratuberculosis antibody enzyme linked immunosorbent assay (ELISA) (IDEXX Inc.) was fully automated using an automation workstation system (AWS). The IDEXX commercial kit is used to screen for exposure to the Johne's disease (JD) bacterium in cattle on a herd basis. Due to the large number of sera involved in this type of surveillance project, there is a need to increase the cost effectiveness and decrease the turn around time. The purpose of the work was to develop an automated ELISA (a-ELISA) for JD antibodies screening and examine the productivity advantages, quality assured performance characteristics, the laboratory information management (LIM) work, and the multitasking provisions under this format. The AWS was assembled using a modular approach design. The high-throughput screening configuration consisted of a Beckman Coulter hybrid Biomek FX liquid-handling system integrated with a Molecular Devices Spectramax microplate reader, a Biotek ELx405 microplate washer, a Kendro Cytomat 6000 hotel, and an Axis PTZ Network camera. The integrated system function was programmed with an IBM computer using the SAMI software. This setup was housed in a Class 2 Biosafety Cabinet. This fully automated system provided flexibility to accommodate the multistep requirements of the immunoassay procedure. Complexity of the working steps was kept to a minimum by using one unique operation program. The complete robotic workstation with its peripheral equipment was simple to operate, robust, and easily maintained. These characteristics enabled the elimination of forced downtime due to human and/or machine failure. A 5 to 10 fold improvement in cost, test capacity, and productivity was achieved. With this system, one diagnostician is able to carry out over 1000 antibody screening tests a day. Based on the automated nature of the test, there is ample walk away time between the ELISA tests, enabling the technologist to multitask–performing other important duties such as sample receiving and preparation, test validation, result interpretation, information dissemination, and other related LIM work. The performance characteristics–such as repeatability, reproducibility, optimal cutoff values, identity score, test sensitivity, and specificity were evaluated. These test validation results were based on data accumulated in an 8-year proficiency testing with the US National Veterinary Services Laboratory. Our laboratory test has received certification from the United States Department of Agriculture to perform serologic testing for JD since 1998. In 2000–2001, this test scope was accredited by the Standard Council of Canada. In 2004, this a-ELISA was validated according to the requirements of the ISO/IEC 17025 standard.
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Comment cette classification a été obtenuedéplier
Prédiction distillée sur la base complète
Imitation des enseignantsNi prévalence calibrée, ni vérité terrain. Validation humaine à venir. Apprise à partir de 10 348 étiquettes directes de Codex et de 10 348 étiquettes directes de Gemma. Le mode candidate est l'union des têtes enseignantes seuillées; le consensus est leur intersection. Ces sorties portent le statut machine_predicted_unvalidated et ne sont ni des étiquettes humaines ni des étiquettes directes de modèles de pointe.
Scores Codex et Gemma par catégorie
| Catégorie | Codex | Gemma |
|---|---|---|
| Métarecherche | 0,003 | 0,003 |
| Méta-épidémiologie (sens strict) | 0,000 | 0,000 |
| Méta-épidémiologie (sens large) | 0,000 | 0,000 |
| Bibliométrie | 0,000 | 0,000 |
| Études des sciences et des technologies | 0,000 | 0,000 |
| Communication savante | 0,000 | 0,001 |
| Science ouverte | 0,000 | 0,000 |
| Intégrité de la recherche | 0,000 | 0,000 |
| Charge utile insuffisante (le modèle a refusé de juger) | 0,000 | 0,000 |
Scores machine (provisoires)
Les deux têtes enseignantes du modèle étudiant, lues sur ce travail. Un score ordonne la base pour la relecture; il n'affirme jamais une catégorie, et le statut de validation accompagne chaque rangée tel quel.
Scores de référence d'un modèle non mature (critères de maturité non atteints, 7 itérations). Un score ordonne; il n'affirme jamais une catégorie.
score_only:v0-immature-baseline · tel quel depuis la passe de notation : score_only signifie que le nombre peut ordonner les travaux, et qu'aucune étiquette de catégorie n'en découleClassification
machine, non validéePrédiction automatique; un appel candidat d’une seule tête enseignante, pas un consensus.
Le détail, modèle par modèle et score par score, se trouve en fin de page sous « Comment cette classification a été obtenue ».